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【目的】对棉铃虫Helicoverpa armigera 2个普通气味受体基因的cDNA全长进行分析,明确这两个普通气味受体基因在不同组织中的表达分布,为进一步的功能研究奠定基础。【方法】利用PCR结合RACE技术克隆棉铃虫两条普通气味受体基因的cDNA全长;利用不同的生物信息学软件对序列进行结构预测、序列比对和进化树分析;利用半定量RT-PCR检测其在棉铃虫成虫不同组织中的表达。【结果】获得两条棉铃虫气味受体基因的全长序列,并命名为HarmOR9和HarmOR29(GenBank登录号分别为KJ188252和KJ188253)。序列分析显示,HarmOR9全长1 206 bp,编码401个氨基酸;HarmOR29全长1 188 bp,编码395个氨基酸。选择已报道的鳞翅目昆虫烟青虫Heliothis assulta、家蚕Bombyx mori、烟芽夜蛾Heliothis virescens和棉铃虫的气味受体与本实验克隆得到的两个气味受体基因的编码产物进行序列比对和进化树分析,结果显示这两个气味受体与性信息素受体区别明显,并与其他普通气味受体聚类在一起。半定量RT-PCR的结果显示HarmOR9与HarmOR29都主要在触角中高表达且无雌雄间差异,HarmOR29在其他组织中均不表达;而HarmOR9在雄虫下唇须中有微量表达,在其他组织中均不表达。【结论】本研究从棉铃虫中克隆得到2个气味受体基因HarmOR9和HarmOR29的cDNA全长,其编码产物具有气味受体的典型特征并且属于普通气味受体。明确了这两个气味受体基因都在棉铃虫成虫的触角中高表达,且无雌雄差异,推测其可能参与了棉铃虫普通气味的识别过程。
【Objective】 The objective of this study was to analyze the cDNA length of two common odorant receptors (Helicoverpa armigera) in Helicoverpa armigera, and determine the expression and distribution of these two common odorant receptors genes in different tissues and lay the foundation for further functional studies. 【Method】 The full-length cDNA of two common odorant receptor genes of Helicoverpa armigera was cloned by PCR combined with RACE technique. The structure, sequence alignment and phylogenetic tree analysis of the sequences were carried out by using different bioinformatics softwares. Semi-quantitative RT-PCR Its expression in different tissues of adult cotton bollworm was detected. 【Result】 The full-length sequences of two odorant receptor genes of H. armigera were obtained and named HarmOR9 and HarmOR29 (GenBank accession numbers KJ188252 and KJ188253, respectively). Sequence analysis showed that the full length of HarmOR9 was 1 206 bp, encoding 401 amino acids. The full length of HarmOR29 was 1 188 bp, encoding 395 amino acids. The reported odorant receptors of Lepidoptera Heliothis assulta, Bombyx mori, Heliothis virescens and Helicoverpa armigera were sequenced and compared with those of the two odorant receptor genes cloned in this experiment Phylogenetic tree analysis showed that the two odorant receptors differed significantly from sex pheromone receptors and clustered with other common odorant receptors. The results of semi-quantitative RT-PCR showed that both HarmOR9 and HarmOR29 were mainly expressed in antennae without any difference between male and female, HarmOR29 was not expressed in other tissues, while HarmOR9 was slightly expressed in the lower lip of male, and in other tissues Do not express. 【Conclusion】 The full-length cDNA of two odorant receptor genes, HarmOR9 and HarmOR29, was cloned from the cotton bollworm in this study. The encoded products have the typical characteristics of odorant receptors and belong to common odorant receptors. It was clear that both of the odorant receptor genes were highly expressed in the antennae of H. armigera adults without any difference between the male and the female, suggesting that they may be involved in the identification of the common odor of H. armigera.